Method for the treatment of hyperglycemia, hyperlipemia, inflammatory diseases, pains or pyrexia due to the accentuation of central nerve, or tumor

ABSTRACT

Disclosed is a method for the treatment of hyperglycemia, hyperlipemia, inflammatory diseases, pains due to the accentuation of central nerve, pyrexia due to the accentuation of central nerve or tumor, which comprises administering to a patient suffering therefrom a pharmaceutically effective amount of p-aminobenzoic acid-N-L-rhamnoside or a pharmaceutically acceptable salt thereof.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a division of application Ser. No. 584,629 filedFeb. 29, 1984, now U.S. Pat. No. 4,555,505, which is acontinuation-in-part of application Ser. No. 484,592 filed Apr. 13,1983, now U.S. Pat. No. 4,559,327 which is a continuation-in-part ofapplication Ser. No. 289,226 filed Aug. 3, 1981, now abandoned, which isa division of application Ser. No. 102,535 filed Dec. 11, 1979, now U.S.Pat. No. 4,313,939, which is a continuation-in-part of application Ser.No. 39,218 filed May 15, 1979, now abandoned.

BACKGROUND OF THE INVENTION

The present invention relates to a method of inhibiting tumor growth ina patient suffering therefrom and a method for the treatment ofhyperglycemia, hyperlipemia, inflammatory diseases, pains due to theaccentuation of central nerve, pyrexia due to the accentuation ofcentral nerve or tumor, which comprises administering to a patientsuffering therefrom a pharmaceutically effective amount ofp-aminobenzoic acid-N-L-rhamnoside or a pharmaceutically acceptable saltthereof.

The inventors of the present invention, during the course of searchingchemical compounds having antitumor activity, have found that theabove-mentioned chemical compound has a number of physiologicalactivities such as blood sugar-reducing activity, antihypertensiveactivity, blood lipid-reducing activity, antiinflammatory activity andcentral nerve-depressing activity in addition to its antitumor activity.

Although the above-mentioned aminobenzoic acid derivatives are knowncompounds, no report has been found on the physiological activity of thecompounds.

"Inoue, et al. N-Glycosides. XIX. Preparation of anthranilic acidN-glycosides., Chemical Abstracts, Vol. 48 (1954), Column 2001 i." and"Inoue, et al. N-Glycosides. XXV. Paper chromatography of N-glycosides.,Chemical Abstracts, Vol. 48 (1954), Column 2003 a." disclose thechemical syntheses of the compounds which are the active ingredients ofthe pharmaceutical composition of the present invention. However, thereis no utility disclosed in these prior arts and no teaching ofpharmaceutical "dosage unit forms".

Furthermore, although U.S. Pat. No. 2,659,689 discloses a p-aldiminobenzoic ester and a composition for protecting the human skin fromerythema producing rale, the composition comprising a solution ofp-aldimino benzoic ester, there is no teaching of pharmaceutical "dosageunit forms".

SUMMARY OF THE INVENTION

In a first aspect of the present invention, there is provided a methodof inhibiting tumor growth in a patient suffering therefrom whichcomprises administering to said patient a pharmaceutically effectiveamount of p-aminobenzoic acid-N-L-rhamnoside or a pharmaceuticallyacceptable salt thereof.

In a second aspect of the present invention, there is provided a methodfor the treatment of hyperglycemia, which comprises administering to apatient suffering therefrom a pharmaceutically effective amount ofp-aminobenzoic acid-N-L-rhamnoside or a pharmaceutically acceptable saltthereof.

In a third aspect of the present invention, there is provided a methodfor the treatment of hyperlipemia, which comprises administering to apatient suffering therefrom a pharmaceutically effective amount ofp-aminobenzoic acid-N-L-rhamnoside or a pharmaceutically acceptable saltthereof.

In a fourth aspect of the present invention, there is provided a methodfor the treatment of inflammatory diseases, which comprisesadministering to a patient suffering therefrom a pharmaceuticallyeffective amount of p-aminobenzoic acid-N-L-rhamnoside or apharmaceutically acceptable salt thereof.

In a fifth aspect of the present invention, there is provided a methodfor the treatment of pains due to the accentuation of central nerve,which comprises administering to a patient suffering therefrom apharmaceutically effective amount of p-aminobenzoic acid-N-L-rhamnosideor a pharmaceutically acceptable salt thereof.

In a sixth aspect of the present invention, there is provided a methodfor the treatment of pyrexia due to the accentuation of central nerve,which comprises administering to a patient suffering therefrom apharmaceutically effective amount of p-aminobenzoic acid-N-L-rhamnosideor a pharmaceutically acceptable salt thereof.

In a seventh aspect of the present invention, there is provided a methodfor the treatment of tumor, which comprises administering to a patientsuffering therefrom a pharmaceutically effective amount ofp-aminobenzoic acid-N-L-rhamnoside or a pharmaceutically acceptable saltthereof.

BRIEF DESCRIPTION OF THE DRAWING

The annexed FIGS. 1 to 2 show respectively the infrared absorptionspectra of respective compound No. 1 to No. 2 in Table 1.

DETAILED DESCRIPTION OF THE INVENTION

The active ingredient of the pharmaceutical composition of the presentinvention is a compound represented by the following formula: ##STR1##wherein ¹ R denotes the residual group formed by removing the OH at1(alpha) or 1(beta) position from rhamnose, or its pharmaceuticallyacceptable salt selected from the group consisting of its Na, K, Mg, Caand Al salts. The sugar moiety of the active ingredient has a structureof a pyranose ring.

The method of preparation of the active ingredient of the presentinvention is illustrated as follows:

A mixture of 4.5 to 5 g of p-aminobenzoic acid, 5-6 g of L-rhamnose and0.1 to 0.5 g of ammonium chloride was heated in 40 to 90 ml of 95 to100% ethanol or pure methanol under a reflux condenser to inducecondensation. After the reaction is over, the reactant is left at roomtemperature or in a cool place and the crystals separated out arecollected by filtering the reactant solution. These crystals are washedwith water, ethanol or ethyl ether, and then recrystallized from anaqueous solution of methanol or ethanol.

In order to substitute the hydrogen atom of the carboxyl group of thethus prepared compound with a base, it is preferable to follow the knownmethod. The compound, paraaminobenzoic acid-N-L-rhamnoside, is dissolvedin an aqueous ethanolic solution and an inorganic salt is added to thesolution to effect the substitution.

The physical properties of the compounds (the active ingredient of thepharmaceutical composition of the present invention) prepared by theabove-mentioned methods are shown in Table 1, and their infraredabsorption spectra are respectively shown in FIGS. 1 to 2. Method ofdetermination of the physical properties are as follows.

                                      TABLE 1    __________________________________________________________________________    Physical properties of the active ingredients                                   Ultraviolet              Melting                     Specific                            Elementary                                   absorption              point  rotation                            analysis (%)                                   Maximum    Compound  (°C.)                     [α] .sub.D .sup.20                            C:H:N  (millimicron)    __________________________________________________________________________    1. p-aminobenzoic              169-170                     -28.6 in                            55.0:6.2:4.8                                   290, 220       acid-N--L-              (decomp.)                     94% ethanol                            (55.1:6.0:4.9)       rhamnoside    2. Sodium p-amino-              173-178                     +80    51.0:5.1:4.8                                   273       benzoate-N--L-              (decomp.)                     in water                            (51.1:5.2:4.6)       rhamnoside    __________________________________________________________________________     Note:     (: : ) = theoretical values of C, H and N (%).

(1) Melting point: determined by the use of micro melting pointdetermination apparatus made by Yanagimoto Works, Japan.

(2) Specific rotation: determination by using direct-reading polarimeterModel OR-50 made by Yanagimoto Works, Japan, with a thickness of 50 mmof an aqueous ethanolic solution of the acidic active ingredient and anaqueous solution of the sodium salt of the acidic active ingredient.

(3) Molecular composition: Elementary analysis was carried out by usingCHN-Coder Model MT-2 made by Yanagimoto Works, Japan.

(4) Ultraviolet absorption spectrum: by using self-recordingspectrophotometer Model PS-3T made by Hitachi Works, Japan, on anaqueous ethanolic solution of the acidic active ingredient and on anaqueous solution of the sodium salt of the acidic active ingredient.

(5) Infrared absorption spectrum: determined by KBr-method usinginfrared absorption spectrometer Model DS-701G made by Nippon Bunko Co.,Ltd., Japan. The chart number of the spectrogram coincides with thenumber of specimens of the active ingredient.

The followings are the physiological properties of the active ingredientof the pharmaceutical composition of the present invention described inthe order of (1) acute toxicity, (2) antimicrobial activity, (3)mutagenicity, (4) delayed-type intracutaneous reaction and (5)antibody-producing activity.

(1) Acute toxicity:

Acute toxicity of sodium p-aminobenzoate-N-L-rhamnoside was examined byrespective intraperitoneal and oral (forcible) administration to ICR-JCLmice. The specimen was dissolved in the physiological saline solution inintraperitoneal administration, and dissolved in distilled water in oraladministration.

Their symptoms were observed after administration until the 7th day ofadministration, and LD₅₀ of the specimen was obtained from the mortalityaccumulated to the 7th day, according to the graphic method ofLitchfield-Wilcoxon. The result is shown in Table 2. As is seen in Table2, sodium p-aminobenzoate-N-L-rhamnoside is qualified to be highly safeactive ingredient of the pharmaceutical composition, having LD₅₀ morethan 10 g/kg, regardless of the route of administration.

                  TABLE 2    ______________________________________    Acute toxicity of the active ingredient                     LD.sub.50 in g/kg                     Route of administration    Compound           Intraperitoneal                                   Oral    ______________________________________    Sodium p-aminobenzoate-N--L-                       15.00       12.80    rhamnoside    ______________________________________

(2) Anti-microbial activity:

Sodium p-aminobenzoate-N-L-rhamnoside was dissolved in distilled waterat a series of two fold dilution system. These diluted solutions weremixed with agar medium in 9 times by volume and the mixture was pouredinto a petridish. Heart infusion agar medium was used for bacteria, andSabouraud's agar medium was used for fungi. After streaking with thepreculture, the inoculated plates were incubated at 37° C. for 20 to 24hours for bacteria and at 25° C. for 3 to 7 days for fungi, and then thegrowth was examined. The following microorganisms were used forassessing the antimicrobial activity:

Pseudomonas aeruginosa IAM 1514

Escherchia coli IFO 12734

Staphylococcus aureus 209 P

Bacillus subtilis IAM 1069

Saccharomyces cerevisiae IAM 4207

Candida albicans ATCC 752

Trichophyton mentagrophytes IFO 6124

Aspergillus niger IAM 3001

As the result of the above-mentioned tests, it was found that sodiump-aminobenzoate-N-L-rhamnoside did not show any growth inhibitionagainst all the microorganisms, at a concentration of 1 mg/ml.

(3) Mutagenicity:

As the first stage, sodium p-aminobenzoate-N-L-rhamnoside was tested byrec-assay (i), and as the second stage, it was tested by reversion assay(ii).

(i) A strain of Bacillus subtilis M 45, a defectant of recombinationrepair, and a wild strain of Bacillus substilis H 17 keepingrecombination repair activity were inoculated to make their own streaksnot crossed at the start on a B-2 agar culture plate (made by dissolving10 g of meat extract, 10 g of polypeptone, 5 g of sodium chloride and 15g of agar in 1000 ml of distilled water at a pH of 7.0). Then, acircular sheet of filter paper 8 mm in diameter, which absorbed 0.04 mlof an aqueous solution of sodium p-aminobenzoate-N-L-rhamnoside (usingsterilized water) was put on the surface of the agar plate so as tocover the starting point of the above-mentioned streaks of bacterialculture. The inoculated B-2 agar culture was kept at 37° C. for a nightand the length of growth-inhibited region was measured. Kanamycin wasused as the negative control and Mitomycin C was used as the positivecontrol. The results of the rec-assay are shown in Table 3.

(ii) The strains TA 98 and TA 100 (both are histidine requiring) ofSalmonella typhimurium were used in the reversion assay.

Into 2 ml of a soft agar culture medium (the medium itself contains 6 gof sodium chloride and 6 g of agar in 1000 ml of distilled water) towhich one tenth by volume of an aqueous solution of 0.5 mM of biotin and0.5 mM of histidine had been added, 0.1 ml of the bacterial suspensionand 0.1 ml of an aqueous solution of sodiump-aminobenzoate-N-L-rhamnoside were admixed and the mixture was layeredon the minimum agar culture medium. After 2 days of incubation at 37°C., the number of revertant colonies was counted. Furylfuramide (AF-2)was used as the positive control. The results of the reversion assay areshown in Table 4.

As is seen in Table 3, sodium p-aminobenzoate-N-L-rhamnoside did notshow any mutagenicity at a high concentration of 5000 microgram/disk.And as is seen in Table 4, the rate of occurrence of mutation by theactive ingredient of the pharmaceutical composition of the presentinvention did not show any difference from that in the control to whichno substance was added, even at a high concentration of 5000microgram/plate. These findings demonstrate that the active ingredientis safe in view of mutagenicity.

                  TABLE 3    ______________________________________    Result of rec-assay             Concentra-                     Length of growth-                                     *Dif-             tion    inhibition zone ference    Compound   (μg/disk)                         H45 (mm)  H17 (mm)                                           (mm)    ______________________________________    Sodium p-amino-               500       0         0       0    benzoate-N--L-               5,000     0         0       0    rhamnoside    Kanamycin  10        5         4       1    Mitomycin C               0.05      12        2       10    ______________________________________     Note:     *Difference = length of inhibition zone of M 45 minus length of inhibitio     zone of H 17.

                  TABLE 4    ______________________________________    Result of reversion assay                  Concen-  Number of revertant                  tration colonies (n/plate)    Compound        (μg/plate)                              TA 100   TA 98    ______________________________________    Sodium p-aminobenzoate-                    5,000      73       4    N--L-rhamnoside    Furylfuramide   0.1       911      167    Control (nothing added)                    --        149      13    ______________________________________

(4) Delayed-type intracutaneous reaction:

In order to know the effects of the active ingredient on cellularimmunity, the foot pad reaction test was carried out using ICR-JCL miceas experimental animals and erythrocytes of sheep as an antigen.

A mouse was primary-sensitized by injecting 0.2 ml of 10% suspension ofsheep erythrocytes in physiological saline solution from the caudal veinand after 7 days of the first sensitization, 0.05 ml of 40% suspensionof sheep erythrocytes in physicological saline solution was injected inthe foot pad for the second sensitization. The thickness of the foot padwas determined on the next day. The administration of the activeingredient of the pharmaceutical composition of the present inventionwas carried out with the dosage of 250 mg/kg/day once a day forconsecutive 5 days centering around the day when the first sensitizationwas carried out.

As the result, the increment of the thickness of the foot pad of themouse administered with the active ingredient showed no significantdifference as compared to the increment in the group of mouse notadministered with the active ingredient.

(5) Antibody-producing activity

In order to know the effects of the active ingredient on humoralimmunity, the hemagglutination test was carried out using ICR-JCL micesensitized with sheep erythrocytes.

A mouse was sensitized by injecting 0.2 ml of 10% suspehsion of sheeperythrocytes in physiological saline solution from the caudal vein andafter 7 days of sensitization the mouse blood was sampled for thehemagglutination test of determination of the autibody-producingactivity. The active ingredient was administered for consecutive 5 dayscentering around the day of sensitization, intraperitoneally at thedosage of 250 mg/kg/day.

As the result, there was no significant difference in agglutinationtiter between the group administered with the active ingredient and thecontrol group.

In the case where the pharmaceutical composition is used as anantiinflammatory agent, it is able to use the pharmaceutical compositionin the form which is convenient to obtain the effectiveness according tothe kinds and the symptoms of the disease, and moreover, the activeingredient may be used as itself or may be used as mixtures combinedwith any diluent allowable in pharmaceutical process and with othermedicines.

The pharmaceutical composition of the present invention is administeredorally or parenterally and accordingly, the pharmaceutical compositionof the present invention may take any form optionally for the oral orparenteral administration.

The pharmaceutical composition of the present invention may be offeredas a form of unit administration. The form of the pharmaceuticalcomposition of the present invention may be powder, granule, tablet,sugar-coated tablet, capsulated one, suppository, suspension, solution,emulsifiable concentrate, ampouled one, injection, etc. As a diluent,any one of solids, liquids and semisolids may be utilized, for instance,excipients, fillers, binders, wetting agents, disintegrating agents,surfactants, demulcents, dispersing agents, buffering agents,surfactants, demulcents, dispersing agents, buffering agents, perfumes,preservatives, dissolution aids and solvents. Moreover, one or more thanone of these adjuvants may be used in combination or in mixtures.

The pharmaceutical composition of the present invention may beformulated by any known method, and the amount of the active ingredientcontained in the composition (preparation) is generally from 0.01% to100% by weight.

The pharmaceutical composition of the present invention may beadministered orally or parenterally to human or animals, however, it ispreferably administered orally. Sublingual administration is included inoral administration. Parenteral administration includes subcutaneousinjection, intramuscular injection and intravenous injection and theinjection by drop method.

The dose of the pharmaceutical composition of the present inventiondepends upon the age, the personal difference and the state of disease,and whether the object is human or animal and accordingly, anextraordinal amount may be administered than the following dose:Generally, for human, the oral dose is 0.1-1000 mg/kg body weight/day,preferably 1-500 mg/kg/day and the parenteral dose is 0.01-200mg/kg/day, preferably 0.1-100 mg/kg/day divided into 1-4 parts, one partbeing administered in one time.

The present invention will be more precisely explained while referringto Examples as follows.

However, the present invention is not restricted to Examples undermentioned. From the foregoing description, one skilled in the art caneasily ascertain the essential characteristics of the present invention,and without departing from the spirit and scope thereof, can makevarious changes and modifications of the invention to adapt it tovarious usages and conditions.

EXAMPLE 1 (Production of p-aminobenzoic acid-N-L-rhamnoside and itssodium salt)

A mixture of 3 g of p-aminobenzoic acid, 4 g of L-rhamnose and 0.1 g ofammonium chloride was heated in 30 ml of 94% ethanol under a refluxcondenser. After the reaction was over, crystals separated out when thereaction mixture was left at room temperature. The crystals thusobtained by filtering were washed with water and an aqueous methanolicsolution, and then recrystallized from 50% methanol. The recrystallizedcrystals were colorless needles. Yield was 30.9%.

Thus obtained p-aminobenzoic acid-N-L-rhamnoside was dissolved graduallyinto an aqueous 1% sodium hydroxide solution containing in total thecalculated amount of sodium hydroxide and after filtering, the solutionwas condensed under reduced pressure. The crystals which separated outby the addition of a large excess of acetone to the condensate wasdehydrated and dried. Colorless crystal of sodium salt was obtained atthe yield of 100% based on the p-aminobenzoic acid-N-L-rhamnoside and atthe total yield of 30.9%.

EXAMPLE 2 (Formulation)

10 parts by weight of sodium p-aminobenzoate-N-L-rhamnoside,

15 parts by weight of (heavy) magnesium oxide and

75 parts by weight of lactose were uniformly mixed and formulated intopowder or granules. The power is filled in capsules to be capsulatedformulation.

EXAMPLE 3 (Formulation)

45 parts by weight of sodium p-aminobenzoate-N-L-rhamnoside,

15 parts by weight of starch,

16 parts by weight of lactose,

21 parts by weight of crystalline cellulose,

3 parts by weight of polyvinyl alcohol and

30 parts by weight of water were uniformly mixed, crushed andformulated, and then dried and shifted to be granules.

EXAMPLE 4 (Formulation)

Granules were prepared as in Example 3, and the mixture of 96 parts byweight of this granule and 4 parts by weight of calcium stearate wascompression-formulated to be tablets 10 mm in diameter.

EXAMPLE 5 (Formulation)

94 parts by weight of sodium p-aminobenzoate-N-L-rhamnoside,

6 parts by weight of polyvinyl alcohol and

30 parts by weight of water were mixed and the mixture was processed asin Example 3 to be granules. To 90 parts by weight of the thus processedgranules 10 parts by weight of crystalline cellulose were mixed and themixture was compression-formulated to be tablets 8 mm in diameter. Thetablets were coated with syrup, gelatine and precipitated calciumcarbonate to be coated tablets.

EXAMPLE 6 (Formulation)

0.6 part by weight of sodium p-aminobenzoate-N-L-rhamnoside,

2.4 parts by weight of a non-ionic surfactant and

97 parts by weight of physiological saline solution were mixed underheating and then the mixture was sterilized to be an injection.

EXAMPLE 7

The followings are the pharmacological properties of the activeingredient of the pharmaceutical composition of the present inventiondescribed in the order of (1) blood sugar-reducing activity, (2)antihypertensive activity, (3) anti-tumor activity, (4) analgeticactivity, (5) antipyretic activity, (6) antiinflammatory activity and(7) blood lipid reducing activity.

(1) Blood sugar-reducing activity

Streptozotocin was administered intraperitoneally to a group of Wistarrats at a dosage of 60 mg/kg and after confirming the positivity ofurinary sugar to the animals on the 8th day, regular insulin was furtheradministered to the rats to reduce both the urinary sugar and the bloodsugar. Out of thus treated animals, those which certainly showed ahigher urinary sugar value and also a higher blood sugar value after afew days of insulin-administration were used as the model animalssuffering from artificial diabetes mellitus. Sodiump-aminobenzoate-N-L-rhamnoside was administered to the model animalsorally as a solution in distilled water at the respective dosages of 30and 300 mg/kg. Blood specimens were collected after 3 and 6 hours of theadministration, and the determination of glucose in the specimen wascarried out by using a RaBA-kit (made by Chugai Pharmaceutical Co.,Japan) according to the enzyme method.

The results are shown in Table 5. As is seen in Table 5, the differencebetween the values of blood sugar before and after the administration ofthe active ingredient (Δ value) was conspicuously larger than Δ value ofcontrol.

                  TABLE 5    ______________________________________    Blood sugar-reducing activity                   Dose Change (Δ value) (mg/dl)                   (mg/ of blood sugar after    Compound         kg)    3 hrs.    6 hrs.    ______________________________________    Sodium p-aminobenzoate-N--                      30    -157      -151    L-rhamnoside     300    -115      -121    Control          --      -36       -39    ______________________________________

(2) Antihypertensive activity

An aqueous solution of sodium p-aminobenzoate-N-L-rhamnoside indistilled water was orally administered to rats of spontaneoushypertension at respective dosages of 30 and 300 mg/kg and their bloodpressure was determined after 3 and 6 hours of administration by asphygmomanometer (made by Ueda Works, Japan, Model USM-105R). Thedifference of blood pressures before and after the administration wasused to evaluate the antihypertensive activity of sodiump-aminobenzoate-N-L-rhamnoside. Mean value of blood pressure of theabove-mentioned rats in spontaneous hypertension was 200 mmHg.

The results are shown in Table 6. As is seen in Table 6, sodiump-aminobenzoate-N-L-rhamnoside clearly showed the antihypertensiveeffect.

                  TABLE 6    ______________________________________    Antihypertensive activity                        Reduced amount of blood                 Dose   pressure after                 rate   3 hrs.    6 hrs.    Compound       (mg/kg)  (mmHg)    ______________________________________    Sodium p-aminobenzoate-                    30      14        14    N--L-rhamnoside                   300      20        25    Control        --       -2*        2    ______________________________________     Note:     *Blood pressure raised by 2 mmHg.

(3) Antitumour activity (against transplanted tumour cells)

(3-1) Against Sarcoma-180:

Sarcoma-180 were transplanted subcutaneously into the right axilla ofICR-JCL mice at the rate of 1×10⁶ cells/mouse, and from after 24 hoursof transplantation an aqueous solution of sodiump-aminobenzoate-N-L-rhamnoside in sterilized physiological salinesolution was orally and forcibly administered every other day at a doserate of 500 mg/kg, 10 times in all. On the 25th day of thetransplantation, the nodular tumour(s) was extirpated and weighed.

The inhibition ratio (I.R.) (%) of the active ingredient was calculatedby the following formula:

    (1-T/C)×100=I.R.(%)

wherein

T: mean weight of the tumour(s) in treated group of mice and

C: mean weight of the tumour(s) in control group* of mice.

The result of the test is shown in Table 7. As is seen in Table 7,sodium p-aminobenzoate-N-L-rhamnoside exhibited a considerably highantitumour activity.

(3-2) Against Melanoma B-16:

A small slice (3 mm in square) of Melanoma B-16 which had beenmaintained by transplantation in generations subcutaneously in C57BL/6mice was transplanted subcutaneously into the axilla of each of femaleBDF₁ mice of 5 weeks after birth (10 animals/group). The specimen,sodium p-aminobenzoate-N-L-rhamnoside, was orally and forciblyadministered daily, in total 10 times, from after 24 hours of thetransplantation at a rate of 100 or 500 mg/kg after dissolving into anaqueous physiological saline solution. By observing the mortality of thethus transplanted mice for 60 days after transplantation, the rate ofelongation of their lives was obtained from the mean survival days andis shown also in Table 7.

(3-3) Against Breast cancer MM-102:

Breast cancer MM-102 which had been maintained by transplantation ingenerations intraperitoneally in C3H/Hc mice and taken from the mouse ofthe 7th generation was intraperitoneally transplanted to each of thefemale mice of the same strain 5 weeks after birth at a rate of 1×10⁶cells/animal (10 animals per group).

The specimen, sodium p-aminobenzoate-N-L-rhamnoside, was orally andforcibly administered daily, in total 10 times, from after 24 hours oftransplantation at a dose rate of 100 or 500 mg/kg after dissolving intoan aqueous physiological saline solution. By observing the mortality ofthe thus transplanted mice for 60 days after transplantation, the rateof elongation of their lives was obtained from the mean survival days,and shown also in Table 7.

(3-4) Against Liver Cancer MH-134:

Liver cancer MH-134 which had been maintained by transplantation ingenerations intraperitoneally in female C3H/Hc mice and taken from themouse of the 7th generation was intraperitoneally transplanted to eachof the female C3H/Hc mice of 5 weeks after birth at a rate of 1×10⁶cells/animal (10 animals per group). The specimen, sodiump-aminobenzoate-N-L-rhamnoside, was orally and forcibly administereddaily, in total 10 times, at 100 or 500 mg/kg after dissolving in anaqueous physiological saline solution to the thus transplanted mice.

By observing the mortality of the thus treated mice for 60 days aftertransplantation, the rate of elongation of their lives was obtained fromthe mean survival days and shown also in Table 7.

                                      TABLE 7    __________________________________________________________________________    Antitumour Activity against Transplanted Tumour    Specimen: Sodium p-aminobenzoate-N--L-rhamnose                     Route of                          Amount of                     Adminis-                          administration                                 Mean survival    Tumour  Animal   tration                          (mg/kg) × 10                                 days    ILS (%)    __________________________________________________________________________    Sarcoma-180            LCR-JCL mouse                     oral 500    --      72.5 (I.R.)*    Malanoma-B-16            BDF.sub.1 mouse                     oral 100    25.6    132.0                          500    26.2    135.1                          0 (control)                                 19.4    --    Breast cancer            C3H/Hc mouse                     oral 100    21.0    129.6    MM-102                500    22.3    137.7                          0 (control)                                 16.2    --    Liver cancer            C3H/Hc mouse                     oral 100    21.7    140.3                          500    21.9    141.3                          0 (control)                                 15.5    --    __________________________________________________________________________     Note:     Against Sarcoma180 only the index of tumour growth inhibition was tested.

(4) Analgetic activity

Determination by the mechanical stimulation method (by applyingpressure)

Female ICR mice which showed a threshold value of pain of 50 to 80 mmHgwhen their tail base part was pressured by a pressure stimulationapparatus (made by Natsume Works, Japan) of Takagi and Kameyama werechosen at test animals, ten animals comprising a group.

After administering sodium p-aminobenzoate-N-L-rhamnoside, the test wascarried out as the time passes and the applied pressure and the timeperiod until the animal showed a quasi-escaping reaction were determinedto evaluate the analgetic activity of the active ingredient.

The result is shown in Table 8. As is seen in Table 8, the pressureapplied on animals when the animal showed the quasi-escaping reactionwas higher in animals to which sodium p-aminobenzoate-N-L-rhamnoside hadbeen applied than in animals not administered, and the time period fromthe beginning to the time point when the animal showed the reaction waslonger in animals administered with the active ingredient than inanimals not administered. Thus, the analgetic activity of sodiump-aminobenzoate-N-L-rhamnoside was confirmed.

Determination by the chemical stimulation method

Sodium p-aminobenzoate-N-L-rhamnoside was orally administered to a group(ten animals) of female ICR mice of age of 5 to 6 weeks, and after 30min of the administration an aqueous 0.6% acetic acid solution wasintraperitoneally injected into the mouse at a dose rate of 0.1 ml/10 gof body weight. The number of writhing syndrome which occurred in themouse during 10 minutes after 10 minutes of intraperitonealadministration was recorded. The analgetic activity was evaluated fromthe writhing syndrome inhibiting ratio obtained by the followingformula:

    (1-T/C)×100=writhing syndrome-inhibiting ratio(%)

wherein

T: mean number of writhing syndrome in the group administered.

C: mean number of writhing syndrome in the control group.

The result is shown in Table 9. As is seen in Table 9, the activeingredient of the pharmaceutical composition of the present inventionshowed analgetic activity. The above-mentioned process was carried outfollowing the method of Kostet et al. (1959).

                  TABLE 8    ______________________________________    Analgetic activity by the mechanical stimulation method                    Quasi-escape reaction pressure                    time until                    (mmHg)   (sec)    Compound          occurrence    ______________________________________    Sodium p-aminobenzoate-N--L-                      92         42    rhamnoside    Control           70         33    ______________________________________     Note:     Amount of administration, 1000 mg/kg p.o.

                  TABLE 9    ______________________________________    Analgetic activity by the chemical stimulation method    Compound                 I.R. (%)    ______________________________________    Sodium p-aminobenzoate-N-- L-rhamnoside                             75.0    ______________________________________     Note:     Amount of administration was 1000 mg/kg p.o.

(5) Antipyretic activity

Following the method of Winter et al. (1961), a 20% suspension of beeryeast was subcutaneously injected to a group (consisting of 6 animals)of rats, and after 10 hours of fasting, sodiump-aminobenzoate-N-L-rhamnoside was orally administered to the rats andtheir rectal temperature was determined.

The antipyretic activity is expressed by the ratio of inhibiting pyrexiadue to beer yeast (I.R. %) at the time when the antipyretic activity ofthe active ingredient is at its maximum according to the followingformula: ##EQU1## wherein T: mean rectal temperature of rats to whichthe active ingredient was administered.

C₁ : mean rectal temperature of rats injected beer yeast, without theactive ingredient.

C₂ : mean rectal temperature of untreated rats (control).

The result is shown in Table 10. As is seen in Table 10, sodiump-aminobenzoate-N-L-rhamnoside exhibited a considerable antipyreticactivity.

                  TABLE 10    ______________________________________    Antipyretic activity                          Antipyretic activity                          (suppressing pyrexia)    Compound              I.R. (%)    ______________________________________    Sodium p-aminobenzoate-N--L-rhamno-                          91.4    side    ______________________________________

(6) Antiinflammatory activity

(a) Carrageenin-edema inhibitory activity

Following the method of Van Arman et al. (1963), sodiump-aminobenzoate-N-L-rhamnoside was forcibly and orally administered toeach rat of a group consisting of 10 animals at the dose rate of 1000mg/kg, and after one hour of the administration 0.1 ml of 1% suspensionof carrageenin in physiological saline solution was injected to theirright foot pad. The volume of the foot pad was determined as time passesand the antiinflammatory activity was expressed by the ratio ofinhibition of the swelling of the foot pad due to carrageenin by theactive ingredient, using the determined value of 1-4 hours from theinjection and calculating by the following formula:

    (1-T/C)×100=I.R.(%)=antiinflammatory activity

wherein

T: mean value of volumes of foot pad in administered animals.

C: mean value of volume of foot pad of control (not administered andthen injected).

The result is shown in Table 11. As is seen in Table 11, the ingredientshowed the inhibitory activity against the edema caused by carrageenin.

(b) Antigranuloma activity

Following the method of Winter et al. (1963), two cotton wool pelletswere implanted into the skin of back of each rat of a group consistingof 6 rats at the symmetrical positions having the median line as theaxis of symmetry, the weight of one pellet being 30±1 mg. Oraladministration of 1000 mg/kg/day of sodiump-aminobenzoate-N-L-rhamnoside was carried out for consecutive 7 days.On the 8th day, the granuloma formed in the rats was extirpated andweighed after drying. The antigranuloma activity expressed by the ratioof inhibition of the growth of the granuloma (I.R., %) was calculated ina manner as shown in (6) (a), and the result is shown in Table 11. As isseen in Table 11, sodium p-aminobenzoate-N-L-rhamnoside exhibited theinhibiting activity of growth of the granuloma.

(c) Antiexudation activity

Following the method of Baris et al. (1965), a volume of air wasinjected subcutaneously in the back of each rat of a group consisting of6 rats to make an air pouch, and then 0.5 ml of 1% cotton oil solutionin sesame oil was injected into the pouch. The oral administration of1000 mg/kg/day of sodium p-aminobenzoate-N-L-rhamnoside was then begunto continue for 5 days. On the 6th day, the amount of exudated liquidinto the pouch was determined and the antiexudation activity expressedby the ratio of inhibitory activity to exudation was calculated in amanner as shown in (6) (a). The result is shown in Table 11. As is seenin Table 11, sodium p-aminobenzoate-N-L-rhamnoside exhibited theantiexudation activity.

(d) Antiadjuvant arthritis activity

Following the method of Fujiwara et al. (1971), Mycobacteriumtuberculosis suspended in liquid paraffin was injected subcutaneouslyinto the right foot pad of each rat of group consisting of 6 rats. After14 days of the injection, rats with similar volume of the foot pad werechosen to form groups (10 animals/group), sodiump-aminobenzoate-N-L-rhamnoside was orally administered daily from the15th day for consecutive 7 days. The volume of the foot pad of rats wasdetermined, and the antiadjuvant arthritis activity of the activeingredient was calculated as the ratio of inhibiting the swelling of thefoot pad by using the formula shown in (6) (a). The result is shown inTable 11. As is seen in Table 11, sodium p-aminobenzoate-N-L-rhamnosideexhibited the antiadjuvant arthritis activity.

                  TABLE 11    ______________________________________    Antiinflammatory activity                            Granu-  Exu-    Compound        Edema   loma    dation                                          Arthritis    ______________________________________    Sodium p-aminobenzoate-                    21.7    22.1    16.2  35.9    N--L-rhamnoside    ______________________________________     Note:     Amount of administration of sodium paminobenzoate-N--Lrhamnoside = 1000     mg/kg.

(7) Blood lipid reducing activity

Japanese male white rabbits were fed for about 3 months with solid feed(CR-1) containing 1% of cholesterol and those animals in which theincrease of seral lipid component was confirmed were used as the modelanimals having experimental arteriosclerosis.

An aqueous solution of sodium p-aminobenzoate-N-L-rhamnoside indistilled water was administered respectively at the dose rates of 30and 300 mg/kg orally and after the administration, blood specimen wascollected as time passes from the auricular vein and the change of totalcholesterol (determined by the enzyme method), phospholipid (determinedby the enzyme method) and beta-lipoprotein (determined by turbidmetry)in the serum was observed.

The results are shown in Table 12. In Table 12, the values of serumcholesterol (mean value of 550 mg/dl), of phospholipid (mean value of320 mg/dl) and of beta-lipoprotein (mean value of 2500 mg/kg) beforeadministration were respectively subtracted from the respective valuesafter 3 and 6 hours of the administration, and only the differences areshown, respectively. Therefore, the minus value shows the decrease andthe plus value shows the increase of the respective values due to theadministration. As is clearly seen in Table 12, sodiump-aminobenzoate-N-L-rhamnoside exhibited the activity of reducing thelipid components as compared to control.

                                      TABLE 12    __________________________________________________________________________    Activity of reducing blood lipids                  Phospholipid                          beta-Lipopro-                                  Cholesterol             Dose (mg/dl) tein (mg/dl)                                  (mg/dl)             (mg/kg)                  3 hrs.                      6 hrs.                          3 hrs.                              6 hrs.                                  3 hrs.                                      6 hrs.    __________________________________________________________________________    Sodium p-amino-              30  -49 -47 -134                              -161                                  -50 -45    benzoate-N--L-             300  -57 -51  -24                              -232                                  -75 -70    rhamnoside    Control         0 -19   0  +3  +8  -4    __________________________________________________________________________

What is claimed is:
 1. A method for the treatment of hyperlipemia, whichcomprises administering to a patient suffering therefrom apharmaceutically effective amount of p-aminobenzoate-N-L-rhamnoside or apharmaceutically acceptable salt thereof.
 2. A method of claim 1 whereinthe active ingredient is said pharmaceutically acceptable salt which isthe sodium salt.
 3. A method of claim 2 via oral administration whereinsaid pharmaceutically effective amount per day is from about 1 to 500 mgper kg.
 4. A method of claim 2 via parenteral administration whereinsaid pharmaceutically effective amount per day is from about 0.01 toabout 200 mg per kg.